PXD015118

PXD015118 is an original dataset announced via ProteomeXchange.

Title	Multiplexed MRM concentration measurements of >200 endogenous proteins in dried blood spots for biomarker discovery and validation
Description	The use of protein biomarkers as surrogates for clinical endpoints requires extensive multilevel validation including development of robust and sensitive assays for precise measurement of protein concentration. Multiple reaction monitoring (MRM), is a well-established methodology that can be used for reproducible protein-concentration measurements in biological specimens collected via microsampling. The dried blood spot (DBS) microsampling technique can be performed non-invasively without the expertise of a phlebotomist, and can enhance analyte stability which facilitate the application of this technique in retrospective studies. Thus, precise, sensitive, and multiplexed methodologies for measuring protein concentrations in DBSs can be used for de novo biomarker discovery. To achieve this goal, MRM assays were developed for multiplexed concentration measurement of proteins in DBS. The lower limit of quantification (LLOQ) was found to have a median total coefficient of variation (CV) of 18% for 245 proteins, while the median LLOQ was 5 fmol of peptide injected on column, and the median inter-day CV over 4 days for measuring endogenous protein concentration was 8%. The majority (88%) of the assays displayed parallelism, while the peptide standards remained stable throughout the assay workflow and after exposure to multiple freeze-thaw cycles. For 190 proteins, the measured protein concentrations remained stable in DBS stored at ambient laboratory temperature for up to 2 months. Finally, the developed assays were used to measure the concentration ranges for 200 proteins in twenty same sex, same race and age matched individuals.
HostingRepository	PanoramaPublic
AnnounceDate	2020-01-06
AnnouncementXML	Submission_2020-01-06_11:33:02.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Azad Eshghi
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Label:13C(6)15N(2); Label:13C(6)15N(4); Label:13C(6)15N(1)
Instrument	6490 Triple Quadrupole LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-21 21:22:21	ID requested
⏵ 1	2020-01-06 11:33:04	announced

Eshghi A, Pistawka AJ, Liu J, Chen M, Sinclair NJT, Hardie DB, Elliott M, Chen L, Newman R, Mohammed Y, Borchers CH, 200 Proteins in Dried Blood Spots for Biomarker Discovery and Validation. Mol Cell Proteomics, 19(3):540-553(2020) [pubmed]

submitter keyword: MRM, DBS, protein, concentration, spike in

Christoph H. Borchers
contact affiliation	University of Victoria - Genome BC Proteomics Centre, University of Victoria, Victoria, BC, Canada
contact email	christoph.borchers@mcgill.ca
lab head
Azad Eshghi
contact affiliation	University of Victoria Proteomics Centre
contact email	azad@proteincentre.com
dataset submitter

Panorama Public dataset URI